Transformation of mammalian cells with an amplifiable dominant-acting gene.
Open Access
- 1 June 1980
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 77 (6) , 3567-3570
- https://doi.org/10.1073/pnas.77.6.3567
Abstract
A mutant hamster gene coding for an altered dihydrofolate reductase was transferred to wild-type cultured mouse cells by using total genomic DNA from methotrexate-resistant Chinese hamster ovary A29 cells as donor. By demonstrating the presence of hamster gene sequences in transformants, direct evidence for gene transfer was provided. Transformants selected for increased resistance to methotrexate contain increased amounts of the newly transferred gene. This mutant dhfr gene was used to introduce the Escherichia coli antibiotic resistance plasmid pBR322 into animal cells. Amplification of the dhfr sequences results in amplification of the pBR322 sequences as well. The use of this gene may allow the introduction and amplification of virtually any genetic element in various new cellular environments.Keywords
This publication has 24 references indexed in Scilit:
- Detection of specific sequences among DNA fragments separated by gel electrophoresisPublished by Elsevier ,2006
- Rabbit β-globin mRNA production in mouse L cells transformed with cloned rabbit β-globin chromosomal DNANature, 1979
- Transformation of mammalian cells with genes from procaryotes and eucaryotesCell, 1979
- Gene Amplification and Drug Resistance in Cultured Murine CellsScience, 1978
- Phenotypic expression in E. coli of a DNA sequence coding for mouse dihydrofolate reductaseNature, 1978
- Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donorCell, 1978
- Biochemical transformation of mouse cells by fragments of herpes simplex virus DNACell, 1977
- Transfer of purified herpes virus thymidine kinase gene to cultured mouse cellsCell, 1977
- Elevated dihydrofolate reductase messenger RNA levels in methotrexate-resistant BHK cellsCell, 1976
- Defective transport of amethopterin (methotrexate) as a mechanism of resistance to the antimetabolite in L5178Y leukemic cellsBiochemical Pharmacology, 1962